Knockout and functional analysis of BSSS-related genes in Acremonium chrysogenum by novel episomal expression vector containing Cas9 and AMA1

被引：6

作者：

Liu, Ling ^{[1
]}

Chen, Zhen ^{[1
]}

Tian, Xiwei ^{[1
]}

Chu, Ju ^{[1
]}

机构：

[1] East China Univ Sci & Technol, State Key Lab Bioreactor Engn, Shanghai 200237, Peoples R China

来源：

BIOTECHNOLOGY LETTERS | 2022年 / 44卷 / 5-6期

关键词：

Acremonium chrysogenum; Cephalosporin C; CRISPR; Cas9; Yeast bud site selection system;

D O I：

10.1007/s10529-022-03255-w

中图分类号：

Q81 [生物工程学（生物技术）]; Q93 [微生物学];

学科分类号：

071005 ; 0836 ; 090102 ; 100705 ;

摘要：

Objective The target sorB gene, related to sorbicillinoid production, and the free expression element, AMA1, were used to verify the methodological approach in Acremonium chrysogenum. Result CRISPR-Cas9 episomal expression system was used to introduce a point mutation into the sorB gene and the addition of sorB donor DNA achieved complete knockout of target genes. Four BSSS (yeast bud site selection system)-related genes, axl1, axl2, bud3, and bud4 were knocked out without impact on yield, dry weight, or pH. Relationships between morphology and stress tolerance in knockout strains were analyzed. Conclusion The gene-editing system used in the current study exceeded 80% efficiency and arthrospores development was found to differ from that in wild-type strain.

引用

页码：755 / 766

页数：12

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